JP6412927B2 - Use of odipalcil in the treatment of mucopolysaccharidosis - Google Patents

Description

The present invention relates to the use of odipalcil or a pharmaceutical composition containing said compound in the treatment of mucopolysaccharidosis, in particular maloto-ramie disease.

Mucopolysaccharidosis (MPS) is a degenerative genetic disease associated with enzyme deficiency. Specifically, MPS is caused by deletion or inactivation of lysosomes, which are enzymes that catalyze the slow metabolism of complex sugar molecules called glycosaminoglycans (GAGs). If the enzyme is deficient in this way, GAG accumulates in the patient's cells and tissues, especially cell lysosomes, causing permanent and progressive cell damage, affecting the patient's appearance, physical ability, organ function, etc. Affects most of the mental development.

Eleven types of enzyme deficiency have been identified corresponding to seven types of MPS. Each MPS is characterized in that one or more kinds of enzymes decomposing mucopolysaccharides, that is, heparan sulfate, dermatan sulfate, chondroitin sulfate and keratan sulfate are deficient or inactivated.

Mucopolysaccharidosis type III (MPS type III) or Sanfiliposis is a lysosomal storage disease characterized by severe and rapid intellectual decline in the mucopolysaccharidosis group. The first symptoms appear at 2-6 years of age and show signs of problem behavior (hyperactivity, aggression), reduced intelligence and sleep disorders, and moderate morphological abnormalities. Nerve damage becomes more prominent around the age of 10, and there is a loss of psychomotor acquisition and communication with the surroundings. After 10 years of age, epilepsy occurs frequently. This disease occurs because one of the four enzymes required to catabolize heparan sulfate is missing and its undegraded product is present. MPS type III is a type IIIA due to the difference in the defective enzyme. (Heparance sulfamidase), type IIIB (α-N-acetylglucosaminidase), type IIIC (acetyl CoA: α-glucosaminide-N-acetyl-transferase) and type IIID (N-acetylglucosamine-6-sulfate sulfatase) Divided into subtypes. At present, there is no effective treatment for this disease.

Mucopolysaccharidosis type VI (MPS type VI), or maloto-ramie disease, is a lysosomal storage disease in the mucopolysaccharidosis group that shows severe physical symptoms but no mental or intellectual regression. . This mucopolysaccharidosis is a rare disease, and the incidence is about 1 in 250,000 to 600,000 newborns. In severe cases, the first clinical symptoms occur in 6-24 months and get worse. Symptoms include facial dysmorphism (glossy tongue, always half-open mouth, thick features), joint contractures, very severe dysplasias (spondylosis, kyphosis, Scoliosis, pigeon chest, hallux valgus, long bone curvature), short stature (less than 1.10 m), liver hypertrophy, valvular heart disease, cardiomyopathy, hearing loss, corneal opacity, and the like. Usually, intelligence development is normal or normal in appearance, but hearing and visual damage can cause learning disabilities. Symptoms and severity in this disease vary considerably from patient to patient, with intermediate and very mild symptom types (vertebral dysplasia associated with cardiovascular injury). Like other mucopolysaccharidosis, maloto-ramie disease is associated with a deficiency in mucopolysaccharide-metabolizing enzymes (in this case, N-acetylgalactosamine-4-sulfatase (also referred to as arylsulfatase B)). This enzyme metabolizes the sulfate group of dermatan sulfate (Non-patent Document 1). When this enzyme is deficient, slow degradation of dermatan sulfate is inhibited, and dermatan sulfate accumulates in lysosomes of storage tissues. At present, only one type of Naglazyme® (recombinant human galsulfase) has been approved as a drug to treat this disease, but its cost is very high (about 350 for the US). $, 000 / year). An alternative treatment for this is allogeneic bone marrow transplantation.

Mucopolysaccharidosis type VII (MPS VII type) or Sly disease is a lysosomal storage disease that is extremely rare in the mucopolysaccharidosis group. Its overall symptoms are very heterogeneous. Prenatal type (non-immune fetal edema), neonatal severe type (dysmorphia), hernia, hepatosplenomegaly, congenital clubfoot, heteroostosis, marked hypotension, growth delay and (Including neurological symptoms that cause severe intellectual disability) and extreme moderate type (thoracic kyphosis) found in adolescence or adulthood. This disease results from the deficiency of β-D-glucuronidase and causes the accumulation of various glycosaminoglycans (dermatan sulfate, heparan sulfate and chondroitin sulfate) in lysosomes. At present, there is no effective treatment for this disease.

Thus, it is clear that it is necessary to provide drug treatment to MPS type III, type VI and type VII patients, and in the case of MPS type VI, to provide alternative treatments that are not based on biotechnology. .

Neufeld et al. "The mucopolysaccharides", The Metabolic Basis of Inherited Diseases, eds. Scriver et al, New York, McGraw-Hill, 1989, p. 1565-1587

Odiparcyl (4-methyl-2-oxo-2H-1-benzopyran-7-yl-5-thio-β-D-xylopyranoside; CAS137215-12-4) belongs to the thioxyloside family. This compound described in patent application EP0421829A is represented by the following formula.

This compound was the subject of clinical development (phases 1 and 2) in the treatment of thrombosis in the late 1990s and early 2000s. The mechanism of action can be summarized as follows. Odipalcil acts as a substrate for the enzyme GT1 (galactosyltransferase 1) that initiates the synthesis of GAG chains towards the dermatan sulfate / chondroitin sulfate pathway. These GAGs become cellular components as proteoglycans (when bound to proteins via serine and the first sugar, xylose), and are also secreted into the extracellular medium. Its roles range from clotting control (heparin / heparan and dermatan sulfate secreted into the blood circulation) to growth factor regulation (β-glycans).

Odipalcil can increase total GAG synthesis at the extracellular level, and can further reduce the intracellular GAG load by acting as a decoy, making the residual effect of N-acetylgalactosamine-4-sulfatase more effective It has been noted that this can be done and this is the subject of the present invention. Therefore, it is considered that MPS type III, type VI and type VII can be treated by reducing GAG accumulation at the intracellular level.

FIG. 5 shows that odipalcil increases the ³⁵ S-labeled GAG concentration in a culture supernatant of bovine aortic endothelial cells in a dose-dependent manner. FIG. 3 shows that odipalcil stimulates total GAG synthesis by human dermal fibroblasts in a dose-dependent manner (+ 94% at 10 μM).

According to a first aspect, the present invention relates to odipalcil for use in the treatment of mucopolysaccharidosis type III, type VI or type VII, in particular MPS type VI (also referred to as Maroto Lamy disease).

Odipalcil and a method for producing the same are described in, for example, EP0421829A.

In the present invention, “odiparcyl” means “β-D-xylopyranoside” type.

In one embodiment, odipalcil used in the present invention is at least 60%, preferably at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or at least 99% D structure. In the present embodiment, the odipalsil is preferably a β-anomeric type.

In another embodiment, odipalcil used in the present invention is at least 60%, preferably at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or at least 99% in β-anomeric form. Preferably there is.

Odipalcil is preferably administered at a rate of about 100 to about 5000 mg per day. For example, about 100, 250, 300, 375, 400, 500, 750, 800, 1000, 1500, 2000, 3000, 4000 or 5000 mg of odipalcil is administered per day.

In one embodiment, odipalcil is administered at least about 0.1 to about 70 mg per kg body weight of the patient per day. For example, at least about 1 or 2 mg to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70 mg of odipalcil is administered per day per kg of patient weight. .

In one embodiment, odipalcil is administered once or twice per day (eg, every 10-12 hours). Therefore, the above-mentioned daily dose can be divided into twice a day (bid) administration. For example, a daily dose of 1000 mg can be administered in two doses of 500 mg. Each dose may consist of one or more dosage forms. For example, a dose of 500 mg may be converted into two dosage forms of 250 mg each.

In one embodiment, odipalcil is administered in a fasted state (ie, when the stomach is empty, eg, at least 1 hour before a meal or after more than 2 hours after a meal). In another embodiment, odipalcil is administered during meal intake (i.e., at the same time or immediately before eating, e.g., about 20-30 minutes before meal or within 5 minutes after meal).

In one embodiment, odipalcil is pharmaceutically acceptable according to methods well known to those skilled in the art (eg, the method described in “Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, 2006”). Formulated as a pharmaceutical composition containing one or more agents.

That is, according to the second aspect, the present invention relates to odipalcil and one or more pharmaceutically acceptable ones used for the treatment of mucopolysaccharidosis type III, type VI or type VII, in particular for the treatment of Maroto Lamy disease. It is related with the pharmaceutical composition containing the excipient | filler made.

The pharmaceutical composition can be in any dosage form suitable for the desired route of administration. Administration routes include oral, tongue, sublingual, oral, rectal, topical, intravenous, intraarterial, subcutaneous, intranasal, transdermal, intramuscular or intraperitoneal.

In one embodiment, the pharmaceutical composition contains about 100-1000 mg of odipalcil, for example, 100, 125, 150, 250, 375, 400, 500 or 1000 mg.

In one embodiment, the pharmaceutical composition is administered by injection and contains a vehicle. The vehicle is typically a sterile aqueous solution that may contain one or more components such as sugars, preservatives, salts, and buffering agents in addition to water. The injectable suspension may contain a suspending agent and a desired liquid vehicle.

In one embodiment, the pharmaceutical composition is administered orally. Suitable oral dosage forms include solid formulations and solutions. When the pharmaceutical composition is a solid preparation (gelatin capsule, tablet, dry powder, etc.), examples of usable excipients include diluents, lubricants, binders, disintegrants, and fillers. The solid preparation may or may not be coated. If coated, the coating may be enteric or non-enteric. When the pharmaceutical composition is a liquid (such as an elixir or syrup), usable excipients include water, glycol, salt water, alcohol, and fragrance.

The pharmaceutical composition is preferably a tablet. Such a composition can be prepared in one or more steps including mixing the various components until a homogeneous mixture is obtained and compressing the mixture to obtain a tablet. In one embodiment, the composition can be prepared by wet granulation. This method is well known to those skilled in the art. For example, odipalcil is mixed, granulated, dried and pulverized with a diluent, a binder, and a part or the whole of a sufficient amount of granulation fluid (such as water) to form granules. If necessary, the granules are mixed with the remaining components and the mixture is compressed. Preferably, the tablet contains about 5% to about 90% odipalcil relative to the total weight of the tablet.

According to a third aspect, the present invention provides a method of treating mucopolysaccharidosis type III, type VI or type VII, in particular maloto-ramie disease, for a patient in need of administration. It relates to a method comprising administering an effective amount of odipalcil or a pharmaceutical composition containing said compound. In one embodiment, the daily usage of odipalcil and the pharmaceutical composition is as defined above.

Hereinafter, the present invention will be described with reference to examples.

<Pharmacological activity>
(1. Results obtained with cultured cells)
(1.1. Bovine aortic endothelial cells)
Bovine aortic endothelial cells (ECACC 92010601) were cultured in a 6-well plate, and odipalcil solubilized with ³⁵ S sodium sulfate (10 μci / ml) and a predetermined concentration (1 to 10 μM; final DMSO concentration: 0.1%) with DMSO And incubated for 24 hours. Thereafter, the culture supernatant was recovered, the cell layer was washed with phosphate buffer (PBS), and the culture supernatant and washing solution were combined together in a test tube. An unlabeled dermatan sulfate (200 μg) solution was then added to act as an entraining agent. Gel filtration using a Sephadex G25 column removed ³⁵ S that was not introduced. At this time, GAG eluted in the column exclusion fraction (V0). A cetylpyridinium chloride solution (final concentration: 0.1%) was added to the eluent, and GAG was precipitated at room temperature for 24 hours. Thereafter, the sample was centrifuged to remove the supernatant. The obtained precipitate was redissolved in 2M magnesium chloride, and GAG was precipitated with 5 volumes of 95% ethanol. After centrifugation, the alcoholic precipitate was redissolved in 0.9% sodium chloride, scintillation fluid was added to the counting vial, and the radioactivity of the preparative fraction was measured.

In order to classify the GAG produced in the supernatant of the cultured cells, the redissolved alcoholic precipitate was treated at 37 ° C. for 3 hours with 0.5 mU / μL of chondroitinase ABC (Proteus vulgaris). After the enzyme was inactivated at 100 ° C. for 3 minutes, undigested GAG was precipitated overnight at 4 ° C. using 5 volumes of 95% ethanol. After centrifugation, the above alcoholic precipitate was redissolved in 0.9% sodium chloride, scintillation fluid was added to the counting vial, and the radioactivity of the preparative fraction was measured.

Heparan sulfate GAG was treated with 4 mU / μl of heparinase II (Flavobacterium heparinum) at 30 ° C. for 12 hours. After the enzyme was inactivated at 100 ° C. for 3 minutes, undigested GAG was precipitated overnight at 4 ° C. using 5 volumes of 95% ethanol. After centrifugation, the above alcoholic precipitate was redissolved in 0.9% sodium chloride, scintillation fluid was added to the counting vial, and the radioactivity of the preparative fraction was measured.

As can be seen from FIG. 1, odipalcil increases the ³⁵ S-labeled GAG concentration in the culture supernatant of bovine aortic endothelial cells in a dose-dependent manner.

Moreover, it is suggested that GAG synthesized by cultured cells by enzyme digestion is mainly chondroitin sulfate type.

(1.2. Human fibroblasts)
Normal human skin fibroblasts (BIOalternatives PF2) were cultured in 96 well plates for 24 hours. After replacing the medium with a medium containing or not containing (control) medium with a predetermined concentration (1 μM, 3 μM, 10 μM) or medium containing 10 ng / ml TGF-β reference (positive control), To evaluate total GAG synthesis, ³ H-glucosamine radiolabel was added and the cells were incubated for 72 hours. At the end of incubation, chaotropic buffer was added to the wells of the culture plate to lyse fibroblasts. Next, the entire GAG of the cell lysate was purified by ion exchange chromatography (Q-Sepharose column). Radioactivity incorporated into the anionic fraction was measured by liquid scintillation.

As can be seen from FIG. 2, odipalcil stimulates total GAG synthesis by human dermal fibroblasts in a dose-dependent manner (+ 94% at 10 μM). Data were statistically analyzed by one-way analysis of variance and Dunnett's test was performed (* p <0.05 vs control; ** p <0.01 vs control; *** p <0.001 vs control) .

(2. Results obtained by oral administration to rabbits in vivo)
Odipalcil was orally administered to New Zealand rabbits at a dose of 400 mg / kg. Four hours after the administration, the animal was anesthetized, and a blood sample was collected in a citrate tube by catheterization to the carotid artery. After centrifugation, plasma was removed and frozen. Plasma GAG was isolated after digesting the protein with Pronase E for 48 hours at 50 ° C. Trichloroacetic acid was added to precipitate proteins and protein residues and incubated overnight at 4 ° C. After centrifugation, the supernatant was collected and dialyzed against 100 volumes of phosphate buffer at 4 ° C. for 48 hours. Cetylpyridinium chloride solution (final concentration: 0.1%) was added to the dialysate to precipitate GAG for 24 hours at ambient temperature. The sample was then centrifuged and the supernatant was removed. The resulting precipitate was redissolved in 2M sodium chloride, and GAG was precipitated with 5 volumes of 95% ethanol. After centrifugation, the alcoholic precipitate was redissolved in 0.9% sodium chloride and desalted with a Sephadex G25 column (PD10).

The extracted plasma GAG was quantified by analyzing the uronic acid content by a modified carbazole method of Bitter and Muir. The qualitative analysis of the plasma GAG extract was performed by subjecting the disaccharide obtained by enzymatic digestion using chondroitinase ABC derived from Proteus vulgaris and chondroitinase AC derived from Arthrobacter aurescens to HPLC.

From the following table, it can be seen that when the animal is treated with odipalcil at a dose of 400 mg / kg, the plasma GAG concentration (measured via the uronic acid concentration) is increased 5-fold compared to the control animal group. I understand. From a qualitative point of view, for chondroitin type GAG, the galactosamine-6-sulfate component and dermatan sulfate component (chondroitin B) measured via galactosamine-4-sulfate disaccharide (Δdi-4S DS) increased. Yes.

From the above results, odipalcil increased total GAG synthesis (human fibroblasts), increased extracellular chondroitin type GAG concentration (bovine aortic endothelial cells), and increased plasma GAG, especially chondroitin type GAG synthesis. I found out that Since MPS type III, type VI and type VII are characterized by intracellular GAG accumulation, these results indicate that odipalcil can reduce intracellular GAG burden and is therefore beneficial for the treatment of MPS.

(Formulation example)
Tablet ingredients (wt%) obtained by wet granulation
Odipalcil: 90%
Crystalline cellulose (National Drug Collection or European Pharmacopoeia): 7%
Povidone (polyvinylpyrrolidone) (US Pharmacopeia or European Pharmacopeia): 3%
Water (US Pharmacopoeia or European Pharmacopoeia): enough to wet granulate

Claims (9)

Including Ojiparushiru, mucopolysaccharidosis type VI or VII type pharmaceutical composition for the treatment of. Ojiparushiru is used to be administered in an amount of 1 00mg~ 5 000mg per day The pharmaceutical composition according to claim 1. The pharmaceutical composition according to claim 1 or 2 , which is administered together with food. The pharmaceutical composition according to any one of claims 1 to 3 , which is used for treatment of mucopolysaccharidosis type VI ( Maloto Lamy disease ) . Containing one or more pharmaceutically acceptable excipients, the pharmaceutical composition according to any one of claims 1 to 4. The pharmaceutical composition according to claim 5 , comprising 100 mg to 1000 mg of odipalcil. It is an oral dosage form , The pharmaceutical composition of any one of Claims 1-6 . The pharmaceutical composition according to claim 7, which is a solid preparation. The pharmaceutical composition according to claim 8, which is a tablet.

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